Dirofilaria repens infection in a dog imported to Norway

Dirofilaria repens infection was diagnosed in a dog that had been imported to Norway from Hungary three years previously. The dog was a four-year-old castrated male mixed-breed dog and presented for examination of two masses on the right thoracic wall. Fine needle sampling from the subcutaneous nodules and subsequent cytological examination revealed a high number of microfilariae and a pyogranulomatous inflammation. At re-examination approximately 3 weeks later, both masses had apparently disappeared spontaneously, based on both inspection and palpation. However, examination of peripheral blood by a modified Knott’s test revealed a high number of unsheathed microfilariae with mean length of 360 μm and mean width of 6-7 μm, often with the classic umbrella handle appearance of D. repens. Polymerase chain reaction and sequencing confirmed the D. repens diagnosis. Subcutaneous dirofilariosis caused by D. repens is probably the most common cause of human zoonotic dirofilariosis in Europe, but currently is rarely encountered in northern countries such as Norway. However, travelling, import and relocation of dogs have increased, and thus the geographical range of these parasites is likely to increase from traditionally endemic southern regions. Increasing numbers of autochthonous cases of D. repens infections in dogs have been reported in eastern and central Europe. Although infection with D. repens often induces only mild signs or subclinical infections in dogs, they nevertheless represent a reservoir for zoonotic transmission and thus a public health concern, and, in addition, due to the long prepatent period and the high frequency of subclinical infections or infections with unspecific clinical signs, could easily be missed. Lack of experience and expectation of these parasites may mean that infection is underdiagnosed in veterinary clinics in northern countries. Also, predicted climate changes suggest that conditions in some countries where this infection is currently not endemic are likely to become more suitable for development in the intermediate host, and thus the establishment of the infection in new areas.     

Effects of temperature and dietary n-3 and n-6 fatty acids on endocytic processes in isolated rainbow trout (Oncorhynchus mykiss,Walbaum) hepatocytes

Effects of different incubation temperatures (2, 8, 14 and 20°C) and hepatocyte membrane fatty acid composition on the rate of internalization and lysosomal degradation of the ligand, mannosylated albumin, that is taken up by receptor-mediated endocytosis, were investigated in rainbow trout (Oncorhynchus mykiss, Walbaum). The fish were kept at a water temperature ranging from 9 to 14°C and fed pelleted diets coated with either capelin oil (control), EPA/DHA-concentrate (rich in n-3 polyunsaturated fatty acids) or soybean oil (rich in n-6 unsaturated fatty acids) for at least 3 months prior to sampling. The endocytic uptake mediated by the mannose receptor was very efficient at all temperatures studied. Lysosomal degradation, on the other hand, came to a halt below 8°C. The activation energies for uptake and degradation were 54.6 and 164.2 kJ/mol respectively. No negative effects of increased amounts of either n-3 or n-6 fatty acids were observed on the endocytic parameters studied. On the contrary, multivariate analysis indicated a positive relationship between high levels of n-6 fatty acids and low unsaturation index in the phosphatidylcholine (PC) fraction of the hepatocytes and the internalization rate of 2°C, meaning that the rate of receptor-mediated endocytosis may be affected by membrane fatty acid composition.     

Acute stress alters the intestinal lining of Atlantic salmon, Salmo salar L.: An electron microscopical study

Groups of Atlantic salmon (Salmo salar L.) in feeding (guts filled with faeces) or fasted (three days of diet deprivation) states were subjected to 15 minutes of acute stress. Blood samples and intestinal tissue were collected and prepared for chemical and ultrastructural analyses at intervals post stress until 53 h of recovery. Subjecting fish to acute stress led to significant alterations of the ultrastructure of the enterocytes lining the gastrointestinal tract (GI tract). The most notable effect was substantial damage to the intercellular junctional complexes in midgut regions. These effects appeared within the first hour after stress, were maintained for at least 12 h and were more pronounced in fed than fasted fish. In contrast, hindgut was influenced less by stress and damage was rarely observed. Stress also influenced fish intestinal microbiota. Adherent bacteria decreased in both midgut and hindgut of stressed fish, and this was accompanied by a significant increase in the bacterial contents of faeces. It is suggested that this was due to the sloughing off of mucus eliminating existing microflora and allowing remaining bacteria (also pathogenic) in the gut lumen to colonize the surface of the enterocytes. Although blood haematocrit and plasma cortisol increased following stress, the response appeared to be greater in fasted fish. There were also significant differences in carbohydrate metabolism. While liver glycogen stores were depleted in fasted fish following the mobilization of glucose into plasma, liver glycogen was never depleted in fed fish. As a consequence, plasma glucose levels remained high for more than 12 h of recovery. In fed fish, plasma lactate was also higher than in fasted salmon, and the clearance rate appeared slower. Acute stress induced oxidative stress, as measured through plasma malondialdehyde, but the effect was marginal and nonsignificant. This revised version was published online in July 2006 with corrections to the Cover Date.     

Browsing of sallow (Salix caprea L.) and rowan (Sorbus aucuparia L.) in the context of life history strategies: a literature review

Sallow (Salix caprea L.) and rowan (Sorbus aucuparia L.) constitute small proportions of the deciduous tree volume in Scandinavia, but are highly preferred winter forage for moose and red deer, which occur at historically high densities. Thus, a possible decline of these tree species has been indicated. Against this background, we have reviewed the life histories of relevance for browsing, as well as the basic biology and genetics of sallow and rowan. The species show similarities with respect to short lifespan, small size and sympodial growth pattern, which are risk factors in a browsing context. They also have high juvenile growth rate, important for growing quickly out of reach of browsers. Sallow depends strongly on disturbance for establishment and is more demanding with respect to soil and light conditions than rowan, possibly important for the substantially lower abundance of sallow on the Norwegian Forest Inventory plots. Similarly, the relative recruitment of small size classes of sallow is less than for rowan. Although recruitment is reported to be hampered in wintering areas with high moose or red deer densities, the inventory data, however, dating only back to 1994, do not suggest a general decrease in any of the species. Sallow and rowan saplings show low mortality in moose and deer dominated areas and the species can be characterised as rather resilient to browsing. Of more concern is that browsing can constrain the development of mature rowan and sallow trees locally, with possible consequences for associated epiphytic biodiversity.     

Interaction of low density lipoproteins with liver cells in rainbow trout

Liver is the main catabolic tissue for low density lipoprotein in rainbow trout (Gjøen and Berg 1992). We have investigated the interaction of LDL with isolated trout liver cells and liver membranes. ¹²⁵I-TC labelled trout LDL bound to isolated trout liver cells in a time dependent and saturable manner with an apparant K_d of 20.1 g/ml, suggesting the existence of a specific binding site on the surface of these cells. The binding was Ca²⁺ dependent assessed by the 50% reduction obtained by 5 mM EDTA. Saturable binding to isolated trout liver membranes could also be demonstrated, but with lower affinity as compared to intact cells. Degradation of ¹²⁵I-TC-LDL in hepatocytes was also saturable as degradation could be inhibited about 60% by a 100 fold surplus of unlabelled LDL. The rate of degradation increased with temperature up to 20°C. Both cell association (binding + uptake) and degradation were reduced down to 20% of control in the presence of microtubular and lysosomal inhibitors. Hepatic catabolism of trout LDL therefore seems to depend on receptormediated endocytosis, followed by lysosomal degradation.Abbreviations TC tyramine cellobiose - LDL low density lipoproteins - MeLDL methylated low density lipoproteins - VLDL very low density lipoproteins - HDL high density lipoproteins - VTG vitellogenin - EDTA ethylenediamine tetraacetic acid - PBS phospate buffered saline - SDS-PAGE sodium dodecyl sulphatepolyacrylamide gel electrophoresis - DMPC L--phosphatidylcholine-dimyristoyl     

Digestive function and intestinal integrity in Atlantic salmon (Salmo salar) fed kernel meals and protein concentrates made from yellow or narrow-leafed lupins

This study assessed the effects of yellow lupin (Lupinus luteus) and narrow-leafed lupin (L. angustifolius) kernel meals and protein concentrates on the gastrointestinal integrity, capacity for digestive hydrolysis, and digestibility of nutrients in Atlantic salmon. A basal diet (FM) was made from fish meal, wheat, and fish oil. Six additional diets were formulated by replacing 30% of the FM diet with lupin kernel meal made from L. l. cv. Wodjil (LKM), L. a. cv. Belara (BKM), and L. a. cv. Myallie (MKM), lupin protein concentrates made from the same L. l. (LPC) and L. a. cv. M (MPC), or extracted soybean meal (SBM). All diets were extruded. Each diet was fed to three groups of 176 g salmon kept in 1 m² tanks with 5.6 °C saltwater for 3 weeks prior to sampling of blood, intestinal organs, digesta, and faeces. Inclusion of lupin meals in the diets resulted in harder and more condensed feed particles. Ulcer-like lesions were observed in the stomach of fish from all feeding groups, and this was worsened by lupin in the diet, but did not appear to be pellet hardness related. No consistent altered morphology was observed in the distal intestine (DI) of fish fed the FM and lupin diets, while the DI of fish fed SBM showed consistent and typical soybean meal-induced pathomorphological changes. Plasma cholesterol was higher when feeding MKM and LKM than when feeding FM, MPC, and LPC, with intermediate levels when feeding BKM and SBM. Feeding LKM and LPC resulted in a higher weight of the GIT when related to body weight. Trypsin activity and bile acid concentration were generally higher in digesta from the pyloric (PI) and mid (MI) intestine when feeding FM and lupin diets than when feeding SBM, while the opposite was seen for trypsin activity in digesta from DI. There were no effects of diet on leucine aminopeptidase (LAP) and maltase activity in PI and MI, but in DI the activity of these brush border enzymes were significantly lowered when feeding SBM. SBM in the diet resulted in watery faeces and lowered apparent digestibility of lipid, but this was not observed when feeding the lupin diets. To conclude, the tested lupin kernel meals and protein concentrates did not alter the intestinal function in Atlantic salmon when included at 30% of the diet. Dietary lupin was, however, involved in the worsening of ulcer-like gastric lesions.     

Pharmacokinetic and pharmacodynamic properties of metomidate in turbot (Scophthalmus maximus) and halibut (Hippoglossus hippoglossus)

Metomidate was administered to halibut (Hippoglossus hippoglossus) and turbot (Scophthalmus maximus) intravenously at a dose of 3 mg/kg bodyweight, as a bath treatment at a dose of 9 mg/L water for 5 min to study the disposition of metomidate, and as bath treatment (9 mg/L) for 10 min to study the absorption and effect of metomidate on respiration and balance/motor control. Additionally, turbot were given metomidate orally at a dose of 7 mg/kg. The studies were performed in seawater at a temperature of 10.3 +/- 0.4 degrees C (halibut) and 18.0 +/- 0.3 degrees C (turbot). Pharmacokinetic modeling of the data showed that metomidate had shorter elimination half-life and higher plasma concentrations in turbot compared with halibut, both species displaying a rapid uptake, distribution and excretion. Following intravenous administration, the volumes of distribution at steady state (Vd(ss)) were 0.21 L/kg (halibut) and 0.44 L/kg (turbot). Plasma clearances (Cl) were 0.099 L/h.kg in halibut and 0.26 L/h.kg in turbot and the elimination half-lives (t(1/2)lambdaz) were calculated to be 5.8 h and 2.2 h in halibut and turbot, respectively. Mean residence times (MRT) were 2.2 h in halibut and 1.7 h in turbot. Following oral administration, the t(1/2)lambdaz was 3.5 h in turbot. The maximum plasma concentration (Cmax) was 7.8 mg/L in turbot 1 h after administration. The oral bioavailability (F) was calculated to 100% in turbot. Following 5 min bath the maximum plasma concentrations (Cmax), which were observed immediately after end of the bath, were 9.5 mg/L and 13.3 mg/L in halibut and turbot, respectively. Metomidate rapidly immobilized the fish, with respiratory depression, reduced heart rate, and loss of balance/motor control within 1 min (mean). Recovery was slow, with resumed balance/motor control after 26.4 min. Opercular respiration movements were resumed more rapidly with a recorded mean of 1.7 min. Oral administration was demonstrated to be a way of immobilizing fish, for example in large aquariums, without exposing them to unwanted stress.     

Tissue distribution of 14C-diflubenzuron in Atlantic salmon (Salmo salar)

Diflubenzuron is a potent inhibitor of chitin synthesis, with potential use against salmon lice infestations. The absorption, distribution and elimination of the substance in Atlantic salmon was examined after a single, oral dose of 75 mg/kg bodyweight. The kinetic properties were studied by whole-body autoradiography, liquid scintillation counting and thin layer chromatography, using a 14C-labelled isotope of the substance. The drug was poorly absorbed from the intestine, but reached a concentration of more than 4 micrograms/g in the mucus layer of the skin 2 days after administration. If maintained for several days, this concentration is probably sufficient to control all moulting stages of sea lice in Atlantic salmon. The main route of excretion was via the bile.     

Occurrence of races and pathotypes of cucurbit powdery mildew in southeastern Spain

The causal agent of cucurbit powdery mildew in southeastern Spain has been investigated since 1996. Of the 139 single-spore isolates obtained, all were identified asSphaerotheca fusca. Four physiological races (1, 2, 4 and 5) of the pathogen were detected. During the survey, a population shift ofS. fusca was observed: race 1 was progressively replaced by other races. In addition, race coexistence was observed in several cucurbit greenhouses. Four host range patterns or pathotypes were distinguished among the isolates ofS. fusca. All isolates were highly aggressive on melon and especially on zucchini cultivars. No clear relationships between races and pathotypes could be established, although isolates virulent on watermelon were preferentially associated with race 1. These data show an apparent heterogeneity ofS. fusca populations in southeastern Spain that should be analyzed further. http://www.phytoparasitica.org posting Aug. 9, 2002.     

Bacterial Apical Necrosis of Mango in Southern Spain: A Disease Caused by Pseudomonas syringae pv. syringae

ABSTRACT A necrotic bacterial disease of mango trees (Mangifera indica) in Spain affecting buds, leaves, and stems is described for the first time. Necrosis of flower and vegetative buds on commercial trees during winter dormancy was the most destructive symptom of the disease. The apical necrosis is caused by Pseudomonas syringae, which was always isolated from mango trees with disease symptoms. Of 95 bacterial strains isolated from symptomatic tissues and characterized from 1992 to 1997, over 90% were identified as P. syringae pv. syringae. Additional strains were isolated from healthy mango trees, and they were identical to the isolates from diseased tissues. Pathogenicity tests on mango plants showed that P. syringae pv. syringae incited the apical necrosis, but that climatic conditions determined the onset of disease development. Populations of total bacteria and of P. syringae and the number of active ice nuclei were monitored over a 3-year period. The largest populations of P. syringae were associated with cool, wet periods that coincided with the highest disease severity, whereas P. syringae was only occasionally detected on healthy trees. The median effective dose was estimated from infectivity titration assays.